LOCUS       sequence-444196-        6600 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Insect cell expression of human 20S CP proteasome subunit beta 
            type-5 (PSMB5), beta type-6 (PSMB6), and beta type-7 (PSMB7) with 
            C-terminal Twin-Strep-tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6600)
  AUTHORS   Adolf F, Du J, Goodall EA, Walsh RM Jr, Rawson S, von Gronau S, 
            Harper JW, Hanna J, Schulman BA
  TITLE     Visualizing chaperone-mediated multistep assembly of the human 20S 
            proteasome.
  JOURNAL   Nat Struct Mol Biol. 2024 Apr 10. doi: 10.1038/s41594-024-01268-9.
  PUBMED    38600324
REFERENCE   2  (bases 1 to 6600)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat Struct 
            Mol Biol. 2024 Apr 10. doi: 10.1038/s41594-024-01268-9."
COMMENT     Sequence Label: pACEBac1-PSMB5-PSMB6-PSMB7-TEV-2xSTII
FEATURES             Location/Qualifiers
     source          1..6600
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(445..536)
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     polyA_signal    692..826
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        complement(1779..1870)
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     polyA_signal    2026..2160
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        complement(3185..3276)
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     protein_bind    3353..3386
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        join(3457..3462,3463..3479,3480..3485)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             3674..4207
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     mobile_element  4274..4497
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     rep_origin      complement(4632..5220)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4712..4731)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     mobile_element  5410..5575
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     polyA_signal    5727..5861
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(join(6033..6056,6057..6092,6093..6116,
                     6117..6122))
                     /codon_start=1
                     /product="two Strep-Tag(R)II moieties connected by a linker
                     for enhanced binding to Strep-Tactin(R), an engineered form
                     of streptavidin (Schmidt et al., 2013)"
                     /label=Twin-Strep-tag
                     /translation="SAWSHPQFEKGGGSGGGSGGSAWSHPQFEK"
     CDS             complement(6129..6149)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="EDLYFQS"
ORIGIN
        1 ttcagcatcc gattggctgt cacaactctg ggaagacggc cagtggagag ggagtggagc
       61 tccaggttgg aagaaatgag ctgggttgtc atgtctgtgt ctgcagctgt cccagcacca
      121 caacaataaa tattaggaga tatgaagtgt atttttgaac agttcttgtc agcaacaacc
      181 atcccttcag ttgctcttgt atctgctcca agaactatgc catccttata gaccacccca
      241 gcgatggtcg tgccagtttt ccggaccttt ggaagcttgt atcccctctt tgcaaaatcg
      301 gcttccaaga cggcattcct gcggcagtta tcaaaagaga agcctccaac tggtggagca
      361 tacaccgaca cagccgccat gaattccgcg cgcttcggac cgggatccgc gcccgatggt
      421 gggacggtat gaataatccg gaatatttat aggttttttt attacaaaac tgttacgaaa
      481 acagtaaaat acttatttat ttgcgagatg gttatcattt taattatctc catgatctat
      541 taatattccg gagtaggtcg atttaaacat caggcatcat taggtttatt taaatggtta
      601 tgatagttat tgctcagcgg tggcagcagc caactcagct tcctttcggg ctttgttagc
      661 agccggatct tctaggctca agcagtgatc agatccagac atgataagat acattgatga
      721 gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga
      781 tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg
      841 cattcatttt atgtttcagg ttcaggggga ggtgtgggag gttttttaaa gcaagtaaaa
      901 cctctacaaa tgtggtatgg ctgattatga tcctctagta cttctcgaca agcttgtcga
      961 gactgcaggc tctagattcg aaagcggccg cttatcaggc gggtggtaaa gtggcaacgg
     1021 cgaatttggg tatctggtct cccaaaagta cttgccgctc tacccctgac tctgcaatgg
     1081 ctgccaggcg gatcactcct ccactggagc catcccgctc catggccaaa gcgagagcat
     1141 tggcagtgaa ttgcagacac tcttccttgg tcatgccttc ccggtaggta gcatcaacat
     1201 agccatagat gtaggagctc ccggagcctc caatggcaaa ggactgcctt accatcatac
     1261 cccccatagg cactgagtac ccctgccctc cttcttgagg gtcccagcct gcgatgatga
     1321 ttcccgccat caggtcttcc cggtatcggt aacacatctc cttaaagagg ctggctgctg
     1381 tgtggaccag tggaggctca ttcagttcaa tgctgtggaa accgagctgg taggtgacag
     1441 catcagctac tgcctgggta tcagcagctg agcctgagcg acagcagaaa atgcggtcgt
     1501 gaataggtgt cagcttgtca gtcactcgat tggcgatgta ggacccagtg gttgttctgg
     1561 agtccgcccc cagaaccacg cccccgtcaa actgcacggc catgatagtc gtcccagtgg
     1621 aaacttctcg gctttcccag tctggagtga atgcctccgg cccccaagcc ggtgctggcc
     1681 cggctccccg agcagctagt aaggtagccg ccatgaattc cgcgcgcttc ggaccgggat
     1741 ccgcgcccga tggtgggacg gtatgaataa tccggaatat ttataggttt ttttattaca
     1801 aaactgttac gaaaacagta aaatacttat ttatttgcga gatggttatc attttaatta
     1861 tctccatgat ctattaatat tccggagtag gtcgatttaa acgtgcaata gtatccagtt
     1921 tatttaaatg gttatgatag ttattgctca gcggtggcag cagccaactc agcttccttt
     1981 cgggctttgt tagcagccgg atcttctagg ctcaagcagt gatcagatcc agacatgata
     2041 agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt
     2101 tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt
     2161 aacaacaaca attgcattca ttttatgttt caggttcagg gggaggtgtg ggaggttttt
     2221 taaagcaagt aaaacctcta caaatgtggt atggctgatt atgatcctct agtacttctc
     2281 gacaagcttg tcgagactgc aggctctaga ttcgaaagcg gccgcttatc agggggtaga
     2341 gccactatac ttctcatgta gatcagccac attgtcactg gagactcgga tccagccatc
     2401 ctcccgcacg tggtagaggt tgactgcacc tcctgagtag gcatctctgt aggtggcttg
     2461 gtagatggct cgacgggcca gatcataggc ctgctccact tccaggtcat aggaatagcc
     2521 ccgatccatg accccatatg catacacaga gccagaacct acagagaagg tggcccctga
     2581 aatccggttc ccttcactgt ccacgtagta gaggccaggg cctctcttat cccagccaca
     2641 gatcatggtg cccatggaca gccccatgcc tttgtactga tacaccatgt tggcaagcag
     2701 tttggaggca gctgctacag agatgcgttc cttatttcga agctcataga ttcgacattg
     2761 ccgagccaac agccgttccc agaagctgca atccgctgcg cccccagcca tggtgcctag
     2821 caggtatggg ttgatctcta tcaccttctt caccgtctgg gaggcaatgt aagcacccgc
     2881 tgtagccctg gagtcagctg caactatgac tccatggcgg aacttgaagg ccagggtggt
     2941 tgttccatga agcatttcga ttcctggctc ttctgggaca ccccagcctg gcgcggccag
     3001 gctcagacca tcactgagac tccctggacc tagatccagc agatctgcac gacccccaag
     3061 tccgaaaaac ccgcgctggt tcaccggtag cggtctctcc aacacgctcg caagcgccat
     3121 gaattccgcg cgcttcggac cgggatccgc gcccgatggt gggacggtat gaataatccg
     3181 gaatatttat aggttttttt attacaaaac tgttacgaaa acagtaaaat acttatttat
     3241 ttgcgagatg gttatcattt taattatctc catgatctat taatattccg gagtaggtcg
     3301 cgaatcgata ctagtgttta aactcgctac cttaggaccg ttatagttac agataacttc
     3361 gtataatgta tgctatacga agttatggcg acttcgctgc tgcccaaggt tgccgggtga
     3421 cgcacaccgt ggaaacggat gaaggcacga acccagttga cataagcctg ttcggttcgt
     3481 aaactgtaat gcaagtagcg tatgcgctca cgcaactggt ccagaacctt gaccgaacgc
     3541 agcggtggta acggcgcagt ggcggttttc atggcttgtt atgactgttt ttttgtacag
     3601 tctatgcctc gggcatccaa gcagcaagcg cgttacgccg tgggtcgatg tttgatgtta
     3661 tggagcagca acgatgttac gcagcagcaa cgatgttacg cagcagggca gtcgccctaa
     3721 aacaaagtta ggtggctcaa gtatgggcat cattcgcaca tgtaggctcg gccctgacca
     3781 agtcaaatcc atgcgggctg ctcttgatct tttcggtcgt gagttcggag acgtagccac
     3841 ctactcccaa catcagccgg actccgatta cctcgggaac ttgctccgta gtaagacatt
     3901 catcgcgctt gctgccttcg accaagaagc ggttgttggc gctctcgcgg cttacgttct
     3961 gcccaagttt gagcagccgc gtagtgagat ctatatctat gatctcgcag tctccggcga
     4021 gcaccggagg cagggcattg ccaccgcgct catcaatctc ctcaagcatg aggccaacgc
     4081 gcttggtgct tatgtgatct acgtgcaagc agattacggt gacgatcccg cagtggctct
     4141 ctatacaaag ttgggcatac gggaagaagt gatgcacttt gatatcgacc caagtaccgc
     4201 cacctaacaa ttcgttcaag ccgagatcgg cttcccggcc gcggagttgt tcggtaaatt
     4261 gtcacaacgc cgcgaatata gtctttacat gcccttggcc acgcccctct ttaatacgac
     4321 gggcaatttg cacttcagaa aatgaagagt ttgctttagc cataacaaaa gtccagtatg
     4381 ctttttcaca gcataactgg actgatttca gtttacaact attctgtcta gtttaagact
     4441 ttattgtcat agtttagatc tattttgttc agtttaagac tttattgtcc gcccacaccc
     4501 gcttacgcag ggcatctgcg catccacagg aagagcgacc caagtcaatc aggggataac
     4561 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     4621 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     4681 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     4741 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     4801 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     4861 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     4921 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     4981 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     5041 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     5101 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     5161 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     5221 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     5281 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     5341 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttgacccaag
     5401 tcaaccggtt gtgggcggac aaaatagttg ggaactggga ggggtggaaa tggagttttt
     5461 aaggattatt tagggaagag tgacaaaata gatgggaact gggtgtagcg tcgtaagcta
     5521 atacgaaaat taaaaatgac aaaatagttt ggaactagat ttcacttatc tggttccaat
     5581 tagatgggta taccctagga aactaagcta tgtgaaccgt ttatttaaat ggttatgata
     5641 gttattgctc agcggtggca gcagccaact cagcttcctt tcgggctttg ttagcagccg
     5701 gatcttctag gctcaagcag tgatcagatc cagacatgat aagatacatt gatgagtttg
     5761 gacaaaccac aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta
     5821 ttgctttatt tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc
     5881 attttatgtt tcaggttcag ggggaggtgt gggaggtttt ttaaagcaag taaaacctct
     5941 acaaatgtgg tatggctgat tatgatcctc tagtacttct cgacaagctt gtcgagactg
     6001 caggctctag attcgaaagc ggccgcttat cacttttcga attgtgggtg agaccaggca
     6061 ctgcccccgc tacccccgcc agagcctcca cctttctcaa actgtggatg gctccaagca
     6121 ctgtcgacag actgaaagta cagatcctcg gaagtgtcca ttgtttggac tgtttcttcc
     6181 agcacctcaa tctccagagg agtgattttc tcagtgagga ctgcagtagt ccctttctca
     6241 cacctgtacc ggccaagcct ggtccccttc ttgttgggca ctgtgtatgg gcggagaaaa
     6301 tccagcttgt tcttgctgat gacgcagagg tcaatgttgc ttccggagcc caggtcgttg
     6361 aagatgccag ctgcgatggc ttcgctcacc agattcttcg cttcctcctc ctccatgtct
     6421 ggcctaaact tatcttcaaa tacagccatt gctgccaagg agccagaacc catggtgaca
     6481 taaggcaact tatcagttga tccatgagga tagatgctgt agaggtgagg tccagtaaca
     6541 tctactcccc ctaaaactag ggctgcacca atgtaacctt gatacctgaa aagcatctgc
//
